Expression, purification and crystallization of human 5-lipoxygenase-activating protein with leukotriene-biosynthesis inhibitors
- Department of Medicinal Chemistry, Merck Research Laboratories, Rahway, NJ 07065 (United States)
- Department of Infectious Diseases, Merck Research Laboratories, Rahway, NJ 07065 (United States)
- Department of Cardiovascular Diseases, Merck Research Laboratories, Rahway, NJ 07065 (United States)
- Department of Pain Research, Merck Research Laboratories, West Point, PA 19486 (United States)
The expression, purification and crystallization of human 5-lipoxygenase-activating protein in complex with two leukotriene-biosynthesis inhibitors is decribed. The processes that were used to generate diffraction quality crystals are presented in detail. The nuclear membrane protein 5-lipoxygenase-activating protein (FLAP) plays an essential role in leukotriene synthesis. Recombinant full-length human FLAP with a C-terminal hexahistidine tag has been expressed and purified from the cytoplasmic membrane of Escherichia coli. Diffraction-quality crystals of FLAP in complex with leukotriene-synthesis inhibitor MK-591 and with an iodinated analogue of MK-591 have been grown using the sitting-drop vapor-diffusion method. The crystals exhibit tetragonal symmetry (P42{sub 1}2) and diffracted to a resolution limit of 4 Å.
- OSTI ID:
- 22360455
- Journal Information:
- Acta Crystallographica. Section F, Vol. 63, Issue Pt 12; Other Information: PMCID: PMC2344111; PMID: 18084092; PUBLISHER-ID: pu5206; OAI: oai:pubmedcentral.nih.gov:2344111; Copyright (c) International Union of Crystallography 2007; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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